首页 > Biosensis > Biosensis/MOAB-2 Mouse Monoclonal antibody to Amyloid beta peptide (A beta 40/42), purified/M-1586-100/100 µg

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Biosensis/MOAB-2 Mouse Monoclonal antibody to Amyloid beta peptide (A beta 40/42), purified/M-1586-100/100 µg

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DescriptionTheamyloidbetapeptideisderivedfromthecleavageoftheAmyloidprecursorprotein(APP)andvariesinlengthfrom39to43aminoacids.However,theform(s)ofamyloid-betapeptide(Aβ)associatedwiththepathologycharacteristicofAlzheimersdisease(AD)remainsunclear.Inparticular,theneurotoxicityofintraneuronalAβaccumulationisanareaofconsiderableresearchandcontroversyprincipallybecauseantibodiesthoughttobespecificforAβhavebeenshowntoactuallydetectintraneuronalAPPandnotAβexclusively.MOAB-2(mouseIgG2b)isapan-specific,high-titerantibodytoAβresidues1-4asdemonstratedbybiochemicalandimmunohistochemicalanalyses(IHC),andishighlyspecificjusttoamyloidbetapeptide.MOAB-2didnotdetectAPPorAPP-CTFsincellculturemedia/lysates(HEK-APPSweorHEKAPPSwe/BACE1)orinbrainhomogenatesfromtransgenicmiceexpressing5familialAD(FAD)mutation(5xFADmice).UsingIHCon5xFADbraintissue,MOAB-2immunoreactivityco-localizedwithC-terminalantibodiesspecificforAβ40andAβ42.MOAB-2didnotco-localizewitheitherN-orC-terminalantibodiestoAPP.Inaddition,noMOAB-2-immunreactivitywasobservedinthebrainsof5xFAD/BACE-/-mice,althoughsignificantamountsofAPPweredetectedbyN-andC-terminalantibodiestoAPP,aswellasby6E10.Inboth5xFADand3xTgmousebraintissue,MOAB-2co-localizedwithcathepsin-D,aMarkerforacidicorganelles,furtherevidenceforintraneuronalAβ,distinctfromAβassociatedwiththecellmembrane.MOAB-2demonstratedstrongintraneuronalandextra-cellularimmunoreactivityin5xFADand3xTgmousebraintissues.RelatedproductsBiotinylatedMOAB-2antibody,cat#M-1742-50-BOligomericAβELISAKit,cat#BEK-2215-1P/2PBatchNo.SeeproductlabelUnitsize100µgAntigenRecombinanthumanamyloidbetaprotein42(Aβ42):DAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVIAAntigenLocationEpitopemapstoresidues1-4ofhumanamyloidbetapeptide40/42AntigenLength42aminoacidsAntibodyTypeMonoclonalIsotypeIgG2b,lamBDaCloneMOAB-2OtherNamesBeta-APP42;Beta-APP40;Beta-amyloidprotein42;Beta-amyloidprotein40;ABPP;APPI;AmyloidbetaA4protein;MOAB2;MOAB-2;Alzheimer"santibody;AB40;AB42;abetaAccessionP05067A4_HUMAN;ProducedinMouseMolecularWeightWithFormicacidextractionsandstrandardreducedwesternproceduresbeta-amyloidpeptidemigratesbetween3-6kDa(seefigure1).PurityThisproductisaProteinApurifiedmouseIgGbin0.02MPotassiumPhosphate,0.15MSodiumChloride,0.01%sodiumazide,pH7.2.ApplicationsWesternBlotting(WB),Immunohistochemistry(IH),Immunohistochemistry/paraffinembeddedIH(P),Immunoprecipitation(IP),Immunofluorescence(IF),ELISA.AntibodyhasbeentestedinWBusingpurifiedsyntheticbeta-amyloidpreparationsandfromtransgenicmousebrainformicacidextracts(seefigure1).Formicacidextraction/concentrationisrequiredforwesternblotdetectionfromextracts.MOAB-2antibodyisspecificforbeta-amyloidanddoesnotdetectAPP.Suggesteddilutionof1:2000-1:5,000forWB,standardECLdetectionsystems.Tissuesamplesforthedetectionofbeta-amyloidshouldbepreparedasdetailedinK.L.Youmansetal.{JournalofNeuroscienceMethods196(2011)5159}forbestresults.Detectionofbeta-amyloid40/42indirectwesternscanbedifficult;Dot-blotsofpreparedsamplesarerecommendedasdetailedinYoumans.KLetal2012.IRorfluorescentdetectionsystemsnotyettested,theybutareexpectedtoworkwellwithhigherprimaryantibodydilutionsbecauseoftheincreasedsensitivityofthedetectionmethods.SuggesteddilutionsforIHCare1:50-1:1,000.Freshfrozen,4%paraformaldehydefixedfrozen,orformalinfixedparaffinembeddedtissuesareallsuitable.Optimaldilutionsmustbedeterminedbytheenduser.Antigenretrievalisrequiredinfixedtissuesforoptimalstaining.Antibodywastestedon4%paraformaldehyde/0.1%glutaraldehydefixedfrozentissuefrom3xTgand5xFADmice.MOAB-2antibodydetectsintraneuronalandextracellularbeta-amyloidinIHCanddoesnotdetectAPP{YoumansKLetal2012}.Theantibodyalsoreactswitharchivalformalin-fixed,paraffin-embeddedtissuesampleswithantigenHeatInducedEpitopeRetrieval(HIER):RecommendedCitrate,pH6.0bufferforHIER.Signalwasweakwithoutantigenretrieval.Immunoreactivelywasexpressedinintraneural-amyloiddeposition(plaque)inAlzheimersbrain.MoAB-2wasfoundtobeextremelycleanandwithanexcellentsignaltonoiseratiowithnoneuro-cellulardiffusivestaining.InadditionMOAB-2demonstratednosignificantdifferencesinAβdetectionusingparaffinfixed,free-floatingsections{YoumansKLetal2012}.Formicacid(FA)treatmentresultedinoptimaldetectionofbothintraneuronalandextracellularAβcomparedtowithoutFA(incubatedin88%FA8min,YoumansKLetal2012).FreefloatingtissuesectionswerepermeABIlizedinTBScontaining0.25%TritonX-100(TBSX;3×10min),blockedwith3%horseseruminTBSX(3×10min)followedby1%horseseruminTBSX(2×10min)andincubatedwithappropriateprimaryantibodiesdilutedinTBSXcontaining1%horseserumovernight.SeeYoumansKLetal2012forfullIH(P)protocolandmethoddetails.ForIF,suggesteddilutionis1:100-1:500.Theantibodywastestedon4%PFAfixedfrozentissue.FixedtissueswerewashedinTBS(3×10min),thenincubatedin88%FA(8min),andthenpermeabilizedinTBSX(3×10min),andblockedinTBSXcontaining5%bovineserumalbumin(BSA;1hr).SectionsweresubsequentlyincubatedwithappropriateprimaryantibodiesdilutedinTBSXcontaining2%BSAovernightonanoscillatoryrotator.Detectionwasviafluorescentlylabelledabsorbedsecondaryantibodies{YoumansKLetal2012}.ForIP,thesuggesteddilutionis1:200to1:1,000forlabeledbeta-amyloidusingProteinA/Gconjugatedbeadsasthecapturevehicle{YoumansKLetal2012}.InanELISA,adilutionof1:50-1:1000issuggested.TheantibodyhasbeentestedinELISAsonsyntheticbeta-amyloidandtissuehomogenatesfrombeta-amyloid-Tgmice.Biosensisrecommendsoptimaldilutions/concentrationsshouldbedeterminedbytheenduserforallapplications.Dilutionsprovidedareonlymeanttoserveasabasicguide.SpecificityMOAB-2detectspreparationsenrichedinU-,O-,F-Aβ42,andU-Aβ40bydot-blot,andisthusapan-specificAβantibody.However,MOAB-2isselectiveforthemoreneurotoxicAβ42comparedtoAβ40.Indeed,MOAB-2demonstratedatitrationagainstantigenconcentration,anddetectsAβ40at2.5pmolbutU-,O-andFAβb42atantigenconcentrationsaslowas~0.1pmol{Youmans.KLetal2012}.MOAB-2doesnotdetectAPP(Amyloidprecursorprotein).SpeciesAgainstHumanAntibodyAgainstHumanCross-reactivityHuman,Rat,otherspeciesnotyettested.ByDotblot,MOAB-2detectedratAβ40andhumanAβ40,albeitwithlessaffinitythanforAβ42.{Youmans.KLetal2012}BlastURLClickhereFormLyophilized,fromaProteinApurifiedpreparationin0.02MPotassiumPhosphate,0.15MSodiumChloride,0.01%sodiumazide,pH7.2;contains0.01%sodiumazideasapreservative.AppearancedrypowderReconstitutionReconstitutein100µlofsterilewater.Centrifugetoremoveanyinsolublematerial.Finalbufferis0.02MPotassiumPhosphate,0.15MSodiumChloride,0.01%sodiumazide,pH7.2.StorageAfterreconstitutionkeepaliquotsat-20°to-70Cforahigherstability.At4°Ckeepuptooneweek,insulated,protectedfromlight;usesterilemethodsandPipettes.Highlypurifiedglycerol(1:1)maybeaddedforanadditionalstability.Avoidrepetitivefreeze/thawcycles.Keeptightlyclosedwhennotinuseandprotectedfromlight.ExpiryDate12monthsafterpurchaseSpecificReferencesZhu,B.etal.(2017)ER-associateddegradationregulatesAlzheimer"samyloidpathologyandmemoryfunctionbymodulatingγ-secretaseactivity.NatCommun.8(1):1472.Application:IHCHuang,TY.etal.(2017)SORLAattenuatesEphA4signalingandamyloidβinducedneurodegeneration.JExpMed.pii:jem.20171413.[Epubaheadofprint].Application:IHCFelecia,M.etal.(2017)PeripheralInflammation,ApolipoproteinE4,andAmyloid-βInteracttoInduceCognitiveandCerebrovascularDysfunction.ASNNeuro.9(4):1759091417719201.Application:IHC/IFThomas,R.etal.(2016)EpidermalgrowthfactorpreventsAPOE4andamyloid-beta-inducedcognitiveandcerebrovasculardeficitsinfemalemice.ActaNeuropatholCommun.4(1):111Application:IHCKoster,KP.etal.(2016)Epidermalgrowthfactorpreventsoligomericamyloid-βinducedangiogenesisdeficitsinvitro.JCerebBloodFlowMetab.[Epubaheadofprint]Application:IFLöffler,T.etal.(2016)DecreasedPlasmaAβinHyperlipidemicAPPSLTransgenicMiceIsAssociatedwithBBBDysfunction.Front.Neurosci.Application:IFKobro-Flatmoen,A.etal.(2016)Reelin-immunoreactiveneuronsinentorhinalcortexlayerIIselectivelyexpressintracellularamyloidinearlyAlzheimer"sdisease.NeuroBIOLOGyofDisease.93:172-183.Application:IHCTai,LM.etal.(2016)TheroleofAPOEincerebrovasculardysfunction.ActaNeuropathol.131(5):709-23.Application:IFKim,YH.etal.(2015)A3DhumanneuralcellculturesystemformodelingAlzheimer"sdisease.NatProt.10(7):985-1006.Application:WBCondello,C.etal.(2015)MicrogliaconstituteabarrierthatpreventsneurotoxicprotofibrillarAβ42hotspotsaroundplaques.NatCommun.6:6176.Application:IFIulitaMFetal(2014)IntracellularAbetapathologyandearlycognitiveimpairmentsinatransgenicratmodeloverexpressinghumanamyloidprecursorprotein:amultidimensionalstudy.ActaNeuropatholCommun.6:61.Application:IF,IHSmithBRetal(2014)Neuronalinclusionsofalpha-synucleincontributetothepathogenesisofKrabbedisease.JPathol.Apr;235(5):509-21.Application:IFGeneralReferencesTaiLMetal(2016)"TheroleofAPOEincerebrovasculardysfunction."ActaNeuropathol.2016Feb16.[Epubaheadofprint]K.L.Youmansetal(2012)IntraneuronalAbetadetectionin5xFADmicebyanewAbeta-specificantibodyMolNeurodegener.2012Mar16;7(1):8.TaiLMetal(2013)LevelsofsolubleapolipoproteinE/amyloid-β(Aβ)complexarereducedandoligomericAβincreasedwithAPOE4andAlzheimerdiseaseinatransgenicmousemodelandhumansamples.JBiolChem.2013Feb22;288(8):5914-26.K.L.Youmansetal(2011)Amyloid-β42altersapolipoproteinEsolubilityinbrainsofmicewithfivefamilialADmutationsJNeurosciMethods.2011Mar15;196(1):51-9.

Biosensis是一家由生命科学家和商业团队组成的公司，致力于为全球学术界和工业界的研究人员提供支持。该团队在基础和应用生命科学研究方面拥有超过100年的经验，并在生命科学研究和诊断市场的生命科学抗体，试剂和试剂盒的商业化方面拥有50多年的经验。